The Positioning of Ciliary Organelles in Hypotrich Ciliates*†

It is commonly observed in hypotrichs that new ciliary rudiments arise directly from or in close juxtaposition to certain pre-existing ciliary elements. Oral primordia often are initiated near specific cirri, cirral rudiments frequently arise as a result of the disaggregation of certain old cirri, and new dorsal ciliature is formed within pre-existing ciliary rows. In the first 2 situations it has been demonstrated experimentally that neither the old ciliature in question nor the specific cortical site marked by that ciliature is essential for the appearance of the new cirral rudiment. The experimental analysis done thus far suggests that the positions of oral and cirral primordia are determined by interacting gradients established in relation to certain reference points. The nature of the reference points is not fully elucidated; in some cases at least these points appear to be more closely related to topographic features of the cell than to specific pre-existing cortical structures. In the dorsal ciliary rows of Euplotes new ciliary units are formed usually and perhaps invariably in close proximity to old ones, and are generally oriented along the axis of the pre-existing row. The result is a tendency to perpetuate the preexisting row number across cell generations. Changes in row number, however, can occur as a result of occasional formation of new units at right angles to the row, a process that is much enhanced in certain homozygous segregants (basal body deficient). The optimal row number (stability range) as well as the number of ciliary units are under genic control. In addition, the spatial pattern of distribution of ciliary units among rows is invariant in all of the material examined. This pattern is presumed to result from an underlying field whose geometry is independent of both the number of units and the number of rows.     

Invention and Mystery in Hypotrich DNA1

ABSTRACT “The capacity to blunder slightly is the real marvel of DNA. Without this special attribute, we would still be anaerobic bacteria and there would be no music.” Lewis Thomas³ Hypotrichs have evolved extraordinary ways of organizing, manipulating, and replicating the DNA in their micronuclear and macronuclear genomes. Short macronuclear DNA molecules containing single genes are created by excision from chromosomes, accompanied by massive elimination of the germline DNA sequences between genes. Germline genes themselves are interrupted by multiple noncoding segments called internal eliminated segments, or IESs, that divide genes into multiple macronuclear-destined segments, or MDSs. The functional significance of this organization is unknown. Over evolutionary time IESs accumulate mutations rapidly are inserted into or excised from genes, and shift position along DNA molecules. MDSs are ligated to create functional genes when IESs are spliced out of micronuclear DNA during macronuclear development. MDSs in some germline genes are in scrambled disorder and become unscrambled in association with IES elimination. Replication of DNA in the macronucleus is accomplished by organization of replication enzymes and factors into a structure that sweeps through the macronucleus to replicate the many millions of gene-sized DNA molecules. The significance of many of the bizarre DNA phenomena in the evolutionary/functional success of hypotrichs is still unclear.     

Chemical Signaling in Ciliates

ABSTRACT. For long, our knowledge of the biology of ciliate pheromones has long relied solely upon the study of the two structurally unrelated gamones identified in culture filtrates of a Blepharisma species. However, the characterization of a number of polypeptide pheromones secreted by Euplotes raikovi and E. octocarinatus has now established that structural relationships of homology usually link these molecules, which is consistent with the genetic basis of the mating type systems evolved by these species. In this context, our growing appreciation of the conserved and variable elements of the pheromone architecture should foster progress in the understanding of pheromone-receptor interactions and thus, provide important clues into pheromone mechanisms of action.     

Differential Nuclear Coloration in Ciliates

SYNOPSIS. Application of the triple stain of Grosso (methyl green-pyronin-orange G) to the exconjugants of different species of ciliates produces a differential coloration of their nuclei. Both the old macronucleus or its remnants and the micronuclei bind methyl green whereas the newly developing macronucleus (macronuclear anlage) has a selective affinity for the orange G present in the mixture. This finding holds true not only for hypotrichous ciliates, whose macronuclear anlagen are involved in the formation of polytene chromosomes, but also for other ciliates not included in the order Hypotrichida. Such a reaction of the new macronuclei apparently results from the presence of non-histone (acidic) proteins associated with the nucleic acids.
The staining method is fully described to provide a more general application.     

纤毛类原生动物中宿主—共生体系统的研究

目前已经在100多种纤毛虫中观察到细菌、藻类和其他微生物等共生体。对纤毛虫中宿主－共生体系统的研究表明,双小核草履虫中卡巴粒的遗传为细胞质遗传理论提供了例证;含细菌共生体的许多厌氧纤毛虫无线粒体,共生体对宿主代谢有重要作用;尾草履虫－钝状全孢螺菌共生作用中,共生菌感染形式的39kDa、15kDa周质蛋白可分别与IF－3－1、IF－3－2两种单抗反应,其共生体早期感染过程中两种抗原的量发生显著变化,并且共生体生殖形式选择性地合成63kDa蛋白质,该蛋白质可能是与共生作有联系的关键分子;绿草履虫－小球藻共生系统中,共生藻中存在葡糖胺硬性壁是其与草履虫发生共生关系的基本条件,其中,共生藻参与宿主代谢,与宿主形成相互受益的专一性关系,并且藻类共生体的作用可能影响了宿主草履虫基因组有关结构,改变了其基因表达。作者推测,探索共生体对宿主基因结构及其表达产物的影响可能是对纤毛虫中共生作用研究的主要趋势,这对于深入了解真核细胞中宿主－共生体双方的相互作用、物质交流在分子水平上的调控机理、细胞结构与功能的关系等细胞生命活动规律是有意义的。     

Foreign Organelle Retention by Ciliates

SYNOPSIS. Intact algal chloroplasts were found by electron microscopy in the peripheral cytoplasm of 2 ecologically important species of planktonic marine ciliates. The exact origin of the chloroplast is uncertain and the periods of their retention by the protozoa is unknown. The 2 ciliate species may function partially or fully as primary producers. In other ciliates algal eye-spots are retained and may actually be utilized. Ecologic and evolutionary implications of the observations are discussed.     

Ciliates from Antarctic sea ice

Five ciliates, Chlamydonella prostomata nov. sp., Paractedoctema acruosa nov. gen., nov. sp., Urocyclon ovatum nov. gen., nov. sp., Porpostoma grassei (Corliss and Snyder 1986) and Cytharoides balechi Tuffrau 1974, collected from sea ice in the Weddell Sea, Antarctica were morphologically and taxonomically investigated. The new genus Paractedoctema is characterized as: Cyclidiidae with naked snout-like apical end and three well-developed membranelles which are multi-rowed in structure and closely apposed one to another; paroral membrane extending anteriorly to about mid-level of M1; one caudal cilium; and silverline system Cyclidium-like. Since Urocyclon Small and Lynn 1985 is a nomen nudum because no type species has been fixed, we re-establish the genus and give a revised definition: uronematids mostly with inverted pear-shaped body and subequatorially positioned cytostome; apical plate dominant; membranelle 1 highly reduced; and paroral membrane extending anteriorly to about mid-level of membrane 2. Based on this new definition, a new combination is suggested: Homalogastra cymruensis (Pérez-Uz and Hope 1997) comb. nov. (formerly Urocyclon cymruensis Pérez-Uz and Hope 1997). For the well-known genus Chlamydonella, an improved diagnosis has been given according to our observations and the data obtained: Lynchellidae without plasmatic protrusions on ventral side; several to many somatic kineties making no noticeable naked gap between left and right areas; perioral kineties continuous or slightly fragmented with leftmost rows parallel to each other, which are arched transversely; and cytopharyngeal rods (nematodesmata) toothed. Macronucleus usually dimorphic. Regarding the related genus Lynchella Kahl 1933, we suggest that the original diagnosis by Kahl should be maintained. Thus, the genus diagnosis is re-provided: Lynchellidae with plasmatic protrusions on ventral side; several to many somatic kineties making no noticeable naked gap between left and right areas; perioral kineties continuous or fragmented with some rows parallel to each other; cytopharyngeal rods toothed; macronucleus generally dimorphic. In the light of the redefinition, a new combination has been made: Chlamydonella nordica (Jankowski 1968) comb. nov. (formerly Lynchella nordica Jankowski 1968). Accepted: 10 October 1999     

A Simple and Rapid PCR-based Method to Isolate Complete Small Macronuclear Minichromosomes from Hypotrich Ciliates: 5S rDNA and S26 Ribosomal Protein Gene of Oxytricha (Sterkiella) nova

Hypotrich ciliates present a macronuclear genome consisting of gene-sized instead of chromosome-sized DNA molecules. Exploiting this unique eukaryotic genome feature, we introduce, for the first time in ciliates, a rapid and easy PCR method using telomeric primers to isolate small complete macronuclear DNA molecules or minichromosomes. Two presumably abundant macronuclear DNA molecules, containing ribosomal genes, were amplified from the Oxytricha (Sterkiella) nova complete genome after using this method, and then were cloned and sequenced. The 5S rDNA sequence of O. (S.) nova is the third one reported among hypotrich ciliates; its primary and secondary structure is compared with other eukaryotic 5S rRNAs. The ribosomal protein S26 gene is the first one reported among ciliates. This “End-End-PCR” method might be useful to obtain similar gene-sized macronuclear molecules from other hypotrich ciliates, and, therefore, to increase our knowledge on ribosomal genes in these eukaryotic microorganisms.     

Morphometric Variation in the Hypotrich Ciliate Genus Euplotes*†

SYNOPSIS. A quantitative method for the analysis of cirral patterns in the genus Euplotes is developed and contrasted with alternative approaches. Inherent variations in cirral pattern, as they are reflected in frequency distributions of intercirral distances, are ascertained through subclonal analyses of Euplotes harpa samples grown under different conditions. This assessment of the extent of variation within a form having 10 frontoventral cirri provides a basis for a comparative study of other Euplotes of cirrotype 10. These results suggest that all marine cirrotype 10 forms with a single dargyrome have the same configuration of frontoventral and transverse cirri. Similarly, most marine double dargyrome forms of cirrotype 10 have this same cirral pattern, and thus constitute variations upon the same morphometric theme.     

Anaerobiosis and Symbiosis with Bacteria in Free-living Ciliates

SYNOPSIS. Marine, sediment-dwelling ciliates were examined for cytochrome oxidase activity by a cytochemical method and for fine structural details. Species of Plagiopylidae (Trichostomatida), i.e. Plagiopyla frontata, Sonderia vorax and Sonderia sp., and of Heterotrichida, i.e., Parablepharisma pellitum, Parablepharisma sp., Metopus contortus, Metopus vestitus and Caenomorpha capucina ; previously considered to be obligate anaerobes because of their sulfide-containing habitat, do not have cytochrome oxidase activity or mitochondria with cristae or tubuli. The evolutionary origin and significance of anaerobic ciliates is discussed. Most of the anaerobic ciliates harbor a flora of ecto- and endosymbiotic bacteria as demonstrated by transmission and scanning electron micrographs. It is speculated that the bacteria may utilize the metabolic end products of the protozoa for growth and energy yielding processes. These associations are also compared with other, previously described cases of symbiosis involving prokaryotes and protozoa.     

Ciliates as engineers of phototrophic biofilms

1. Phototrophic biofilms consist of a matrix of phototrophs, non‐photosynthetic bacteria and extracellular polymeric substances (EPS) which is spatially structured. Despite widespread exploitation of algae and bacteria within phototrophic biofilms, for example by protozoans, the ‘engineering’ effects of these ciliates on the spatial heterogeneity of phototrophic biofilms are poorly studied. 2. We studied the potential engineering effects of two ciliates, Urostyla sp. and Paramecium bursaria, on the spatial heterogeneity of synthetic multispecies biofilms. Biomass of phototrophic organisms, EPS and bacteria was analysed three dimensionally using confocal laser scanning microscopy. Spatial heterogeneity and cover of the phototrophs, bacteria and EPS were determined at several depths within the biofilm. 3. Ciliate species did not interfere with the overall development of phototrophic microorganisms, because the thickness of the biofilm was equal whether the ciliates were present or not, even though their abundance did affect spatial heterogeneity of biofilm components. When Urostyla was present, it reduced aggregation in EPS and bacteria and increased EPS biovolume. This implies a local facilitating effect of ciliates on photosynthetic activity. Biofilms to which Paramecium was added did not differ from controls in terms of phototrophs, EPS cover and biovolume. Nevertheless, ciliates affected the spatial heterogeneity of these components as phototrophs and EPS became more evenly distributed. 4. This study shows that ecosystem engineering by organisms does not only occur at large spatial scales, as in grasslands and estuaries, but also plays a role at the microscopic scale of biofilms. This effect on spatial heterogeneity was not driven by substantial exploitation of biofilm components, but via the subtle engineering effects of ciliates.     

Entrapment of Ciliates at the Water-Air Interface

The importance of water-air interfaces (WAI) on microorganism activities has been recognized by many researchers. In this paper, we report a novel phenomenon: the entrapment of ciliates Tetrahymena at the WAI. We first characterized the behavior of cells at the interface and showed that the cells'' swimming velocity was considerably reduced at the WAI. To verify the possible causes of the entrapment, we investigated the effects of positive chemotaxis for oxygen, negative geotaxis and surface properties. Even though the taxes were still effective, the entrapment phenomenon was not dependent on the physiological conditions, but was instead affected by the physical properties at the interface. This knowledge is useful for a better understanding of the physiology of microorganisms at interfaces in nature and in industry.     

Exocellular Carbohydrase Formation by Rumen Holotrich Ciliates

SYNOPSIS Exocellular carbohydrase activity was detected, in the absence of cell lysis, in cell-free culture supernatant fluids of rumen holotrich ciliates after incubations in buffer systems of varying tonicity, from cell suspensions that were isolated by various technics, and in which bacterial activity had been suppressed by antibiotics. The kinetic characteristics of the holotrich invertases and β–glucosidase from Dasytricha , although having interspecies variations, were the same for the intra- and extracellular form of the enzyme. The properties of the invertase activity present in cell-free rumen contents resembled those of the exocellular enzymes formed by the holotrichs. Invertase activity in the in vitro culture supernatant fluid of Dasytricha ruminantium increased throughout the incubation period and was influenced by the initial pH, temperature, sucrose concentration, and inoculum size. Exocellular holotrich carbohydrase activity was increased when the incubation substrate was not readily utilized by the protozoa. Intracellular carbohydrase activity was also influenced by the carbohydrate substrate.